Investigating How Alcohol Withdrawal Dysregulates 'Reward' and 'Aversion' Neurons in the Basolateral Amygdala

研究酒精戒断如何调节基底外侧杏仁核中的“奖励”和“厌恶”神经元

• 批准号: 10536418
• 负责人: Michaela Price
• 金额: $ 4.68万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10536418
• 关键词: Abstinence; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcohols; Amygdaloid structure; Anti-Anxiety Agents; Anxiety; Area; Automobile Driving; Behavioral; Benzodiazepines; Brain region; Cells; Chronic; Confocal Microscopy; Cues; Data; Development; Distal; Electrophysiology (science); Emotional; Ethanol; External Capsule; Female; Fright; Glutamates; Goals; Heavy Drinking; Immunohistochemistry; Interneurons; Label; Laboratories; Lateral; Learning; Medial; Mediating; Neurons; Nucleus Accumbens; Patients; Pharmacotherapy; Positioning Attribute; Positive Reinforcements; Prefrontal Cortex; Rattus; Relapse; Research; Rewards; Role; Sensory Process; Stress; Structure; Structure of terminal stria nuclei of preoptic region; Synapses; Synaptic plasticity; System; Technical Expertise; Technology; Testing; Training; Withdrawal; Withdrawal Symptom; Woman; alcohol abuse therapy; alcohol exposure; alcohol use disorder; anxiety-like behavior; base; behavior test; behavioral response; career; designer receptors exclusively activated by designer drugs; experimental study; gamma-Aminobutyric Acid; high reward; hippocampal pyramidal neuron; improved; innovation; male; men; neurophysiology; patch clamp; postsynaptic; recruit; sex; stria terminalis; therapeutic target; transmission process; withdrawal-induced anxiety

The basolateral amygdala (BLA) is critical to the development of alcohol use disorder (AUD). Its unique position within reward- and aversion-related circuitry allows it to regulate alcohol intake and withdrawal-induced anxiety. Glutamatergic pyramidal neurons drive these behavioral responses such that higher pyramidal neuron excitability facilitates anxiety and reward-seeking. Pyramidal neurons receive glutamatergic inputs from midline structures like the medial prefrontal cortex through the stria terminalis (ST) and more lateral cortical areas send glutamatergic inputs through the external capsule (EC). GABAergic lateral paracapsular cells (LPCs) and ‘local’ interneurons regulate pyramidal neurons. The goal of my project is to examine how chronic intermittent ethanol and withdrawal (CIE/WD) dysregulates BLA neurons projecting to reward and aversion-related regions to understand the mechanisms driving increased alcohol intake. The nucleus accumbens (NAC) is involved in reward-seeking while the bed nucleus of the stria terminalis (BNST) regulates anxiety. Therefore, my project will use BLA-NAC neurons as ‘reward’ neurons and BLA-BNST neurons as ‘aversion’ neurons. Our laboratory has shown that WD increases glutamatergic function through pre- and postsynaptic mechanisms at ST inputs and EC-BLA synapses, respectively. My preliminary data reveal that WD increases glutamate release from ST inputs onto BLA-BNST neurons regardless of sex, whereas the WD-mediated increase in postsynaptic function at EC-BLA synapses is both projection- and sex-dependent. My preliminary data also indicate that WD alters the excitability of BLA-NAC and BLA-BNST neurons when GABAergic transmission is blocked. In BLA-NAC neurons, females have higher basal excitability and only males increase excitability after CIE/WD. In BLA- BNST neurons, WD increases excitability, regardless of sex. Blocking glutamatergic and GABAergic transmission abolished the increase in excitability, emphasizing the role of glutamatergic transmission in WD- mediated hyperexcitability. This proposal will continue to investigate how WD impacts ‘reward’ and ‘aversion’ BLA neurons. Specific Aim 1 will employ electrophysiology and retrograde labeling to examine GABAergic function in synapses with BLA-NAC and BLA-BNST neurons. Laboratory data reveal that WD suppresses LPC GABA release and postsynaptic GABAergic function in ‘local’ interneuron synapses. Moreover, reduced LPC GABA release is unique to males. Specific Aim 1 will also examine structural plasticity in GABAergic synapses with BLA-NAC and BLA-BNST neurons using immunohistochemistry, confocal microscopy, and retrograde labeling. Specific Aim 2 will combine behavioral approaches and chemogenetics to inhibit specific BLA circuits and evaluate their role in post-CIE alcohol intake. These experiments will establish that ‘reward’ and ‘aversion’ BLA neurons undergo distinct neurophysiological and structural changes, leading to increased alcohol intake after CIE/WD. Identifying the mechanisms driving increased alcohol intake could improve treatments for AUD.

基底外侧杏仁核（BLA）对于酒精使用障碍（AUD）的发展至关重要，其独特之处在于。 在奖赏和厌恶相关电路中的位置使其能够调节酒精摄入和戒断引起的 谷氨酸能锥体神经元驱动这些行为反应，使得更高的锥体神经元。 兴奋性促进焦虑和寻求奖励。锥体神经元接收来自中线的谷氨酸输入。 通过终纹 (ST) 的内侧前额叶皮层和更多外侧皮层区域等结构发送 通过外囊 (EC) 的谷氨酸能输入和 GABA 能外侧囊旁细胞 (LPC) 和 “局部”中间神经元调节锥体神经元，我的项目的目标是研究慢性间歇性如何。 乙醇和戒断 (CIE/WD) 失调投射到奖赏和厌恶相关区域的 BLA 神经元 了解伏隔核 (NAC) 参与的驱动酒精摄入量增加的机制。 寻求奖励，而终纹床核（BNST）调节焦虑。因此，我的项目。 我们的实验室将使用 BLA-NAC 神经元作为“奖励”神经元，使用 BLA-BNST 神经元作为“厌恶”神经元。 已表明 WD 通过 ST 输入的突触前和突触后机制增强谷氨酸能功能 我的初步数据显示，WD 分别增加了 ST 的谷氨酸释放。 无论性别如何，BLA-BNST 神经元的输入，而 WD 介导的突触后功能增加 EC-BLA 突触既依赖于投射，也依赖于性别。我的初步数据也表明 WD 会改变。 当 BLA-NAC 中 GABA 传递受阻时，BLA-NAC 和 BLA-BNST 神经元的兴奋性。 在 BLA- 中，雌性神经元的基础兴奋性较高，只有雄性在 CIE/WD 后兴奋性增加。 BNST 神经元，WD 增加兴奋性，无论性别如何。 传输消除了兴奋性的增加，强调了谷氨酸传输在WD-中的作用 该提案将继续研究 WD 如何影响“奖励”和“厌恶”。 BLA 神经元。具体目标 1 将采用电生理学和逆行标记来检查 GABA 能。 实验室数据显示 WD 抑制 LPC 的突触功能。 “局部”中间神经元突触中的 GABA 释放和突触后 GABA 功能，LPC 减少。 GABA 释放是男性独有的。特定目标 1 还将检查 GABA 能突触的结构可塑性。 使用免疫组织化学、共聚焦显微镜和逆行法与 BLA-NAC 和 BLA-BNST 神经元 具体目标 2 将结合行为方法和化学遗传学来抑制特定的 BLA 回路。 并评估它们在 CIE 后酒精摄入中的作用，这些实验将确定“奖励”和“厌恶”。 BLA 神经元经历明显的神经生理和结构变化，导致酒精摄入量增加 确定导致酒精摄入量增加的机制可以改善 AUD 的治疗。